Morphological assessment of grafted rat and mouse cortical neurons: A light and electron microscopic study

Abstract

The morphology of cortical neurons grafted into (or near) the rat striatum was studied by means of intracellular Lucifer yellow injections in fixed slices. Rat donor syngeneic cortical tissue (from postnatal day 1 old rats; AO strain) as well as mouse donor xenogeneic cortical tissue (prenatal day 19; C3H/HE strain) were grafted as solid pieces into 8-12 week-old rats (AO strain). Recipients of mouse xenografts were immunosuppressed with a monoclonal antibody against the interleukin-2 receptor. After perfusion and sectioning of the graft-containing areas, individual slices were incubated in the DNA stain 4.6-diamidino-2-phenylindole (DAPI) to visualize the cell nuclei. Grafts could be easily identified by a surrounding rim of astrocytes which outline the border between grafted and host tissue. Grafted cortical neurons were intracellularly filled with Lucifer yellow, DAB-photoconverted, and further processed for light and electron microscopy. In general, no cortical lamination could be observed in the grafted rat and mouse cortical tissue, but neurons were loosely packed throughout the graft. Two major cell types could be identified in all grafts investigated so far. The majority resembled those described as spiny neurons (85%), which could be further classified into pyramid-like, spiny stellate-like or fusiform spiny neurons, with somata ranging between 15 and 25 microns in diameter. The remaining 15% resembled non-spiny neurons with either a multipolar basket-like or fusiform morphology. Dendrites of spiny and non-spiny neurons, which could extend to distances up to 400 microns, were never seen to cross the astrocytic border, but some main axon and axonal collaterals of spiny neurons were found to leave the graft. On the basis of light microscopic observations no difference was found between mouse and rat grafted cortical neurons. The results of this study show that grafted cortical neurons retain some of the characteristic features of neurons in the intact adult cerebral cortex, although there appears to be a greater preponderance of spiny neurons in grafted tissue. This may reflect an immaturity of the grafted tissue or a response to the striatal environment.